Haynes Lab:Notebook/CRISPR Editing/2016/04/12

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04/12/2016

transfected Luc14s with dCas9 g025, 31, and 34 using Neon, plated into 10cm plates

Neon settings
3mL buffer E2 into Neon tube
Pulse voltage 1,100 V
Pulse width 20
Pulse # 2

Collect cells from flasks, need 5x10^6 cells / transfection
Resuspended into 350ul Resuspension buffer R (ended up with too much volume, should have used less buffer?)
mixed with 20ug DNA, used 100ul tips, plated immediately after transfection
changes for next time

use less resuspension buffer, or somehow modify so the final volume is right



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